Compounds for treating ILK-mediated diseases

ABSTRACT

The present invention relates to compounds that are modulators of Integrin Linked Kinase (ILK), and methods of treating diseases with such compounds. In certain embodiments, the compounds are within Formulas I-VII (e.g., Csbl-1). In some embodiments, the compounds are used to treat an ILK-mediated disease, such as cancer (e.g., triple negative breast cancer) or an inflammatory disease.

The present application is a § 371 national entry of PCT/US2019/014500,filed Jan. 22, 2019, which claims priority to U.S. Provisionalapplication Ser. No. 62/621,941 filed Jan. 25, 2018, each of which isherein incorporated by reference in its entirety.

STATEMENT REGARDING FEDERAL FUNDING

This invention was made with government support under grant numberHL058758 awarded by the National Institutes of Health. The governmenthas certain rights in the invention.

FIELD OF THE INVENTION

The present invention relates to compounds that are modulators ofIntegrin Linked Kinase (ILK), and methods of treating diseases with suchcompounds. In certain embodiments, the compounds are within FormulasI-VII (e.g., Csbl-1). In some embodiments, the compounds are used totreat an ILK-mediated disease, such as cancer (e.g., triple negativebreast cancer) or an inflammatory disease.

BACKGROUND OF THE INVENTION

Breast cancer is the most common cancer for women worldwide, with nearly1.7 million new cases diagnosed and more than half million deaths in2012. Clinically, based on the expression levels of estrogen receptor(ER), progesterone receptor (PR), and human epidermal growth factorreceptor 2 (HER2), breast cancer is classified into subgroups of hormonereceptor-positive, HER-2-positive, and triple-negative breast cancer.Triple-negative breast cancer (TNBC), characterized by the absence ofER/PR and lack of overexpression of HER2, represents approximately15-20% of all breast cancers.

As TNBC does not respond to either hormonal therapy or anti-HER2 agents,standard chemotherapy is currently the mainstay of systemic medicaltreatment therefor. TNBC initially responds to conventionalchemotherapy, however patients frequently have rapid relapses andcurrently there is no effective treatment thereafter. In addition, TNBCis more aggressive than other subtypes of breast cancer, which has greatpropensity to metastasize to the lungs and brain. So patients with TNBCusually have a poor prognosis and a shorter overall survival chancecompared with other subtypes of breast cancer.

SUMMARY OF THE INVENTION

The present invention relates to compounds that are modulators ofIntegrin Linked Kinase (ILK), and methods of treating diseases with suchcompounds. In certain embodiments, the compounds are within FormulasI-VII (e.g., Csbl-1). In some embodiments, the compounds are used totreat an ILK-mediated disease, such as cancer (e.g., triple negativebreast cancer) or an inflammatory disease.

In some embodiments, provided herein are compositions comprising: acompound having a structure of Formula I, or a salt thereof, whereinFormula I is:

wherein R1-R7, R13-R16, and X1 are each an atom or group independentlyselected from: hydrogen, deuterium, methyl, trifluoromethyl, loweralkyl, methoxy, lower alkoxy, aryloxy, trifluoromethoxy, —SCF₃, cyano,nitro, amino, lower alkylamino, lower dialkylamino, mercapto, loweralkylthio, arylthio, formyl, acetyl, lower alkylcarbonyl, arylcarbonyl,lower alkylcarboxy, arylcarboxy, lower alkoxylcarboxy, aryloxylcarboxy,formamido, lower alkanoylamino, arylcarbonylamino, carbamido, loweralkylcarbamido, arylcarbamido, aminocarboxy, lower alkylaminocarboxy,arylaminocarboxy, trifluoroacetyl, halogen, hydroxylcarbonyl, loweralkoxylcarbonyl, aryloxycarbonyl, sulfinyl, lower alkylsulfinyl,arylsulfinyl, sulfonyl, lower alkylsulfonyl, arylsulfonyl, sulfonamido,lower alkylsulfonamido, arylsulfonamido, and aryl;

wherein at least one of R1-R7, R14-R19, and X1 is not hydrogen;

wherein R17-R19 are each independently selected from: hydrogen,deuterium, methyl, trifluoromethyl, lower alkyl, methoxy, lower alkoxy,aryloxy, trifluoromethoxy, —SCF₃, cyano, nitro, amino, lower alkylamino,lower dialkylamino, mercapto, lower alkylthio, arylthio, acetyl, loweralkylcarbonyl, arylcarbonyl, formyl, lower alkylcarboxy, arylcarboxy,lower alkoxylcarboxy, aryloxylcarboxy, formamido, lower alkanoylamino,arylcarbonylamino, carbamido, lower alkylcarbamido, arylcarbamido,aminocarboxy, lower alkylaminocarboxy, arylaminocarboxy,trifluoroacetyl, halogen, hydroxylcarbonyl, lower alkoxylcarbonyl,aryloxycarbonyl, sulfinyl, lower alkylsulfinyl, arylsulfinyl, sulfonyl,lower alkylsulfonyl, arylsulfonyl, sulfonamido, lower alkylsulfonamido,arylsulfonamido, aryl, —OH, —CH₂—OH, —CH₂—NH₂, —CH₂—NO₂, —CH₂—COOH,—CH₂—CN, and —CH₂—CONH₂;

wherein R8-R12 are each independently selected from: hydrogen,deuterium, fluorine, chlorine, bromine, and iodine;

wherein X2 is N or CH;

wherein X3 is NH, O, S, or CH₂; and

wherein X4 is selected from: fluorine, chlorine, bromine, iodine, —CN,and —C≡CY where Y is hydrogen, or deuterium.

In certain embodiments, provided herein are composition comprising: acompound having a structure of Formula II, or a salt thereof, whereinFormula II is:

wherein R1-R14, and R20-R23 are each an atom or group independentlyselected from: hydrogen, deuterium, methyl, lower alkyl, methoxy, loweralkoxy, aryloxy, trifluoromethyl, trifluoromethoxy, —SCF₃, cyano, nitro,amino, lower alkylamino, lower dialkylamino, mercapto, lower alkylthio,arylthio, formyl, acetyl, lower alkylcarbonyl, arylcarbonyl, loweralkylcarboxy, arylcarboxy, lower alkoxylcarboxy, aryloxylcarboxy,formamido, lower alkanoylamino, arylcarbonylamino, carbamido, loweralkylcarbamido, arylcarbamido, aminocarboxy, lower alkylaminocarboxy,arylaminocarboxy, trifluoroacetyl, halogen, hydroxylcarbonyl, loweralkoxylcarbonyl, aryloxycarbonyl, sulfinyl, lower alkylsulfinyl,arylsulfinyl, sulfonyl, lower alkylsulfonyl, arylsulfonyl, sulfonamido,lower alkylsulfonamido, arylsulfonamido, and aryl, and

wherein at least one among R1-R14, R21-R26 is not hydrogen;

wherein R24-R26 are each independently selected from: hydrogen,deuterium, methyl, lower alkyl, methoxy, lower alkoxy, aryloxy,trifluoromethyl, trifluoromethoxy, —SCF₃, cyano, nitro, amino, loweralkylamino, lower dialkylamino, mercapto, lower alkylthio, arylthio,formyl, acetyl, lower alkylcarbonyl, arylcarbonyl, lower alkylcarboxy,arylcarboxy, lower alkoxylcarboxy, aryloxylcarboxy, formamido, loweralkanoylamino, arylcarbonylamino, carbamido, lower alkylcarbamido,arylcarbamido, aminocarboxy, lower alkylaminocarboxy, arylaminocarboxy,trifluoroacetyl, halogen, hydroxylcarbonyl, lower alkoxylcarbonyl,aryloxycarbonyl, sulfinyl, lower alkylsulfinyl, arylsulfinyl, sulfonyl,lower alkylsulfonyl, arylsulfonyl, sulfonamido, lower alkylsulfonamido,arylsulfonamido, aryl, —OH, —CH₂—OH, —CH₂—NH₂, —CH₂—NO₂, —CH₂—COOH,—CH₂—CN, and —CH₂—CONH₂;

wherein R15-R19 are each independently selected from: hydrogen,deuterium, fluorine, chlorine, bromine, and iodine;

wherein X1 is nitrogen or CH;

wherein X2 is selected from: NH, O, S, and CH₂; and wherein X3 isselected from: fluorine, chlorine, bromine, iodine, —CN, and —C≡CY whereY is hydrogen or deuterium.

In other embodiments, provided herein are compositions comprising: acompound having a structure of Formula III, or a salt thereof, whereinFormula III is:

wherein X1 is n-propylene, and wherein 1 to 6 hydrogen atoms areoptionally replaced in said n-propylene with deuterium atoms;

wherein X2 is selected from: —OH, —OD, —OCH₃, —OCH₂D, —OCHD₂, and —OCD₃,wherein D is deuterium;

wherein X3 is N or CH;

wherein X4 is selected from: NH, O, S, and CH₂;

X5 is selected from: fluorine, chlorine, bromine, iodine, —CN, and —C≡CYwhere Y is hydrogen or deuterium;

wherein R1-R8 are each independently selected from: hydrogen anddeuterium;

wherein R9-R13 are each independently selected from: hydrogen,deuterium, fluorine, chlorine, bromine, and iodine;

wherein R14-R17 are each an atom or group independently selected from:hydrogen, deuterium, methyl, lower alkyl, methoxy, lower alkoxy,aryloxy, trifluoromethyl, trifluoromethoxy, —SCF₃, cyano, nitro, amino,lower alkylamino, lower dialkylamino, mercapto, lower alkylthio,arylthio, formyl, acetyl, lower alkylcarbonyl, arylcarbonyl, loweralkylcarbonyl, arylcarbonyl, lower alkylcarboxy, arylcarboxy, loweralkoxylcarboxy, aryloxylcarboxy, formamido, lower alkanoylamino,arylcarbonylamino, carbamido, lower alkylcarbamido, arylcarbamido,aminocarboxy, lower alkylaminocarboxy, arylaminocarboxy,trifluoroacetyl, halogen, hydroxylcarbonyl, lower alkoxylcarbonyl,aryloxycarbonyl, sulfinyl, lower alkylsulfinyl, arylsulfinyl, sulfonyl,lower alkylsulfonyl, arylsulfonyl, sulfonamido, lower alkylsulfonamido,arylsulfonamido, and aryl,

wherein at least one among R14-R20 is not hydrogen; and

wherein R18-R20 are each independently selected from: hydrogen,deuterium, methyl, lower alkyl, methoxy, lower alkoxy, aryloxy,trifluoromethyl, trifluoromethoxy, —SCF₃, cyano, nitro, amino, loweralkylamino, lower dialkylamino, mercapto, lower alkylthio, arylthio,formyl, acetyl, lower alkylcarbonyl, arylcarbonyl, lower alkylcarbonyl,arylcarbonyl, lower alkylcarboxy, arylcarboxy, lower alkoxylcarboxy,aryloxylcarboxy, formamido, lower alkanoylamino, arylcarbonylamino,carbamido, lower alkylcarbamido, arylcarbamido, aminocarboxy, loweralkylaminocarboxy, arylaminocarboxy, trifluoroacetyl, halogen,hydroxylcarbonyl, lower alkoxylcarbonyl, aryloxycarbonyl, sulfinyl,lower alkylsulfinyl, arylsulfinyl, sulfonyl, lower alkylsulfonyl,arylsulfonyl, sulfonamido, lower alkylsulfonamido, arylsulfonamido,aryl, —OH, —CH₂—OH, —CH₂—NH₂, —CH₂—NO₂, —CH₂—COOH, —CH₂—CN, and—CH₂—CONH₂.

In some embodiments, provided herein are compositions comprising: acompound having a structure of Formula IV, or a salt thereof, whereinFormula IV is:

wherein R1 is selected from: 2-methoxyethoxy, 2-ethoxyethoxy,2-morpholinoethoxy, ethynyl, 2-(2-hydroxyethoxy)ethoxy, —O—CH₂CH₂CH₂OH,—O—CH₂CH₂—NO₂, —O—CH₂CH₂COOH, —O—CH₂CH₂—CONH₂, —O—CH₂CH₂—O—CH₂OH,—O—CH₂CH₂—O—CH₂CN, —O—CH₂CH₂—O—CH₂NH₂, —O—CH₂CH₂—O—CH₂NO₂,—O—CH₂CH₂—O—CH₂COOH, —O—CH₂CH₂—O—CH₂CF₃, and —O—CH₂CH₂—O—CH₂CONH₂;

wherein R2 is selected from: hydrogen, methoxy, or methoxyethoxy;

wherein R3 is selected from: —CH₃ and —CH₂CH₂—O—CH₃;

wherein X1 is selected from: N, and CH; and

wherein X2 is selected from: NH, O, S, and CH₂.

In particular embodiments, provided herein are compositions comprising:a compound having a structure of Formula V, or a salt thereof, whereinFormula V is:

wherein R1 is selected from: 2-methoxyethoxy, 2-ethoxyethoxy,2-morpholinoethoxy, ethynyl, 2-(2-hydroxyethoxy)ethoxy, —O—CH₂CH₂CH₂OH,—O—CH₂CH₂—NO₂, —O—CH₂CH₂COOH, —O—CH₂CH₂—CONH₂, —O—CH₂CH₂—O—CH₂OH,—O—CH₂CH₂—O—CH₂CN, —O—CH₂CH₂—O—CH₂NH₂, —O—CH₂CH₂—O—CH₂NO₂,—O—CH₂CH₂—O—CH₂COOH, —O—CH₂CH₂—O—CH₂CF₃, and —O—CH₂CH₂—O—CH₂CONH₂;

wherein R2 is selected from: hydrogen, methoxy, and methoxyethoxy;

wherein R3 is selected from: hydrogen, methoxy, and methoxyethoxy;

wherein X1 is N or CH; and

wherein X2 is selected from: NH, O, S, and CH₂.

In additional embodiments, provided herein are compositions comprising:a compound having a structure of Formula VI, or a salt thereof, whereinFormula VI is:

wherein R1 is selected from: 2-methoxyethoxy, 2-ethoxyethoxy,2-morpholinoethoxy, ethynyl, 2-(2-hydroxyethoxy)ethoxy, —O—CH₂CH₂CH₂OH,—O—CH₂CH₂—NO₂, —O—CH₂CH₂COOH, —O—CH₂CH₂—CONH₂, —O—CH₂CH₂—O—CH₂OH,—O—CH₂CH₂—O—CH₂CN, —O—CH₂CH₂—O—CH₂NH₂, —O—CH₂CH₂—O—CH₂NO₂,—O—CH₂CH₂—O—CH₂COOH, —O—CH₂CH₂—O—CH₂CF₃, and —O—CH₂CH₂—O—CH₂CONH₂;

wherein X1 is N or CH; and

wherein X2 is selected from: NH, O, S, and CH₂.

In certain embodiments, provided herein are compositions comprising: acompound having a structure of Formula VII, or a salt thereof, whereinFormula VII is:

wherein R1 is selected from: 2-methoxyethoxy, 2-ethoxyethoxy,2-morpholinoethoxy, ethynyl, 2-(2-hydroxyethoxy)ethoxy, —O—CH₂CH₂CH₂OH,—O—CH₂CH₂—NO₂, —O—CH₂CH₂COOH, —O—CH₂CH₂—CONH₂, —O—CH₂CH₂—O—CH₂OH,—O—CH₂CH₂—O—CH₂CN, —O—CH₂CH₂—O—CH₂NH₂, —O—CH₂CH₂—O—CH₂NO₂,—O—CH₂CH₂—O—CH₂COOH, —O—CH₂CH₂—O—CH₂CF₃, and —O—CH₂CH₂—O—CH₂CONH₂;

wherein X1 and Y1 are O; and

wherein X2 and Y2 are each independently O or CH₂.

In some embodiments, the compound blocks Mg-ATP binding to humanIntegrin Linked Kinase (ILK). In other embodiments, the compoundmodulates ILK-mediated cytoskeleton assembly. In further embodiments,the compound is an anti-cancer agent. In additional embodiments, thecompound is an anti-inflammatory compound. In further embodiments, thecompound is selected from the group consisting of: Compound 1 (Csbl-1),Compound 3 (cmpd #3), Compound 4 (cmpd #4), Compound 1 new (cmpd #1new),Compound 2 new (cmpd #2new), Compound 7 (cmpd #7), and Compound 8(Csbl-2).

In particular embodiments, provided herein are methods comprising:treating a subject with any of the compounds described above or herein(e.g., administering or providing the compound to a subject). Inparticular embodiments, the subject (e.g., human subject) has at leastone condition selected from the group consisting of: cancer, diabetes,breast cancer, triple negative breast cancer, solid tumors, aninflammatory disease, ulcerative colitis, psoriasis, scleroderma,systemic lupus erythematosus and atopic dermatitis, and inflammatoryarthritis, diabetes, type 2 diabetes, a cardiovascular disease,hypertension, heart attack. In some embodiments, the treating is underconditions such that at least one symptom of said at least one conditionis reduced or eliminated. In further embodiments, the subject is a human(e.g., human female or human male). In certain embodiments, the subjecthas an ILK-mediated disease or condition selected from the groupconsisting of: cancer (e.g., leukemia, colon cancer, lung cancer,prostate cancer, pancreatic cancer, brain cancer, kidney cancer, breastcancer, skin cancer or myeloma), diabetes, thrombosis, aging, heartdisease, and pulmonary disease (e.g., pulmonary hypertension).

In certain embodiments, the compound is co-administered with ananti-cancer agent and/or an anti-inflammatory agent. In particularembodiments, the subject is selected from the group consisting of: i) asubject having a disease, ii) a subject displaying signs or symptoms orpathology indicative of said disease, iii) a subject suspected of havingsaid disease, iv) a subject suspected of displaying signs or symptoms orpathology indicative of said disease, v) a subject at risk of saiddisease, vi) a subject at risk of displaying pathology indicative ofsaid disease, vii) an animal model of said disease, and viii) a healthysubject wishing to reduce risk of said disease.

In particular embodiments, the treating comprises administering between0.05 mg-3000 mg of said compound (e.g., daily) to said subject (e.g.,0.05 . . . 1.0 . . . 150 . . . 500 . . . 2000 . . . or 3000 mg). Infurther embodiments, the treating comprises administering between 25mg-600 mg of said compound (e.g., daily) to said subject (e.g., 25 . . .100 . . . 300 . . . 400 . . . 600 mg). In some embodiments, the treatingcomprises administering 25 mg-600 mg to said subject per day for atleast two days (e.g., for at least 2 . . . 14 . . . 28 . . . 150 . . .or 360 days).

DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawings will be provided by the Office upon request and paymentof the necessary fee.

FIG. 1 , panels A and B, show a hypothetical structural comparison ofthe drug binding modes for ILK and EGFR. (A) Structural comparison ofILK (cyan)-erlotinib (green) and EGFR (brown)-erlotinib (yellow)complexes showing the similar drug binding modes with differentsurrounding residues such as EGFR F778 vs ILK S204. (B) Structuralcomparison of ILK (green)-erlotinib (orange) complex with ILK(blue)-gefitinib (pink) complex showing that the surrounding L207, K220(hydrophobic part), and L267 interact differently with erlotinib andgefitinib.

FIG. 2 , panels A and B, show hypothetical binding of Csbl-1(compound 1) to ILK. Panel A shows the crystal structures of ILK boundto ATP (red) and Csbl-1 (yellow) showing that Csbl-1 sterically occludesATP. Panel B shows a structural comparison of ILK bound to Csbl-1 withEGFR showing that Csbl-1 would exert steric clashes with EGFR such asF778 in the EGFR Gly-rich loop region, whereas the correspondingposition is much smaller sized S204 in ILK.

FIG. 3 shows the off target analysis of Csbl-1 vs known kinaseinhibitors. Higher selectivity score means stronger off target effects.

FIG. 4 , panels A and B, show the effect of Csbl-1 (compound 1) onIPP-induced actin bundles. Panel A shows IPP-induced actin bundles(Mg2+; ATP=80 μM; 40 μM). Panel B shows increased numbers of smallersizes as a result of Csbl-1 (Csbl-1: Mg2+; ATP=120 μM; 40 μM).

FIG. 5 , panels A and B, show the effects of Csbl-1 and Erlotinib oncells. Panel A shows proliferation of normal breast MCF10a cells. Cellswere seeded at 20,000 cells/ml in 12 well plates in complete media. Onday 3, the compounds were added at final 2 μM and the cells were countedevery 3 days. Panel B shows proliferation of triple negative breastcancer MDA-MB-231 cells. Cells were seeded at 10,000 cells/ml in 12 wellplates in complete media. The compounds were added at final 2 μM on day2 and the cells were counted every 2 days.

FIG. 6 shows the Kaplan-Meier (KM, http:// followed bykmplot.com/analysis/) plot, which reveals the correlation of survival of186 TNBC patients with ILK (Gene ID 201234 at) expression levels. Thehigher the ILK expression, the lower the patient survival.

FIG. 7 , panels A and B, show the effects of Csbl-1 (Compound 1) on TNBCin a xenograft model. Panel A shows results where 4 mice with TNBCtumors were treated by Csbl-1 at 100 mg/kg orally, five times per weekstarting at day 10. As compared to 4 control mice, TNBC tumors weresubstantially reduced after being treated by Csbl-1 with approximately40% reduction in size at day 45. Panel B shows a tumor comparison at day50 with and without Csbl-1 treatment.

FIG. 8 , panels A-C, show the effect of Csbl-1 on TNBC PDX mice. Csbl-1was dissolved in olive oil. Five mice with TNBC were treated orally withCsbl-1 at 200 mg/kg five times/week vs 4 control mice without thetreatment. Panel A shows tumors with the treatment by Csbl-1 (C1) weresignificantly reduced at day 27 vs the control mice. Panel B shows therepresentative tumors in control mice vs panel C where tumors weresubstantially reduced after the Csbl-1 treatment.

DEFINITIONS

As used herein, the terms “host,” “subject” and “patient” refer to anyanimal, including but not limited to, human and non-human animals (e.g.,dogs, cats, cows, horses, sheep, poultry, fish, crustaceans, etc.) thatis studied, analyzed, tested, diagnosed or treated. As used herein, theterms “host,” “subject” and “patient” are used interchangeably, unlessindicated otherwise.

As used herein, the terms “administration,” and “administering,” referto the act of giving a drug, prodrug, or other agent, or therapeutictreatment (e.g., compositions of the present invention) to a subject(e.g., a subject or in vivo, in vitro, or ex vivo cells, tissues, andorgans). Exemplary routes of administration to the human body can bethrough the eyes (ophthalmic), mouth (oral), skin (topical ortransdermal), nose (nasal), lungs (inhalant), oral mucosa (buccal), ear,rectal, vaginal, by injection (e.g., intravenously, subcutaneously,intratumorally, intraperitoneally, etc.) and the like.

As used herein, the terms “co-administration” and “co-administering”refer to the administration of at least two agent(s) (e.g., compositioncomprising a compound of Formula I-VII, and one or more otheragents—e.g., an anti-cancer or anti-inflammatory agent). In someembodiments, the co-administration of two or more agents or therapies isconcurrent. In other embodiments, a first agent/therapy is administeredprior to a second agent/therapy. Those of skill in the art understandthat the formulations and/or routes of administration of the variousagents or therapies used may vary. The appropriate dosage forco-administration can be readily determined by one skilled in the art.In some embodiments, when agents or therapies are co-administered, therespective agents or therapies are administered at lower dosages thanappropriate for their administration alone. Thus, co-administration isespecially desirable in embodiments where the co-administration of theagents or therapies lowers the requisite dosage of a potentially harmful(e.g., toxic) agent(s), and/or when co-administration of two or moreagents results in sensitization of a subject to beneficial effects ofone of the agents via co-administration of the other agent.

As used herein, the term “at risk for disease” refers to a subject(e.g., a human) that is predisposed to experiencing a particulardisease. This predisposition may be genetic (e.g., a particular genetictendency to experience the disease, such as heritable disorders), or dueto other factors (e.g., age, weight, environmental conditions, exposuresto detrimental compounds present in the environment, etc.). Thus, it isnot intended that the present invention be limited to any particularrisk, nor is it intended that the present invention be limited to anyparticular disease.

As used herein, the term “suffering from disease” refers to a subject(e.g., a human) that is experiencing a particular disease. It is notintended that the present invention be limited to any particular signsor symptoms, nor disease. Thus, it is intended that the presentinvention encompass subjects that are experiencing any range of disease(e.g., from sub-clinical manifestation to full-blown disease) whereinthe subject exhibits at least some of the indicia (e.g., signs andsymptoms) associated with the particular disease.

As used herein, the term “pharmaceutical composition” refers to thecombination of an active agent (e.g., composition comprising a compoundof Formulas I-VII) with a carrier, inert or active, making thecomposition especially suitable for diagnostic or therapeutic use invitro, in vivo or ex vivo.

The terms “pharmaceutically acceptable” or “pharmacologicallyacceptable,” as used herein, refer to compositions that do notsubstantially produce adverse reactions, e.g., toxic, allergic, orimmunological reactions, when administered to a subject.

As used herein, the term “topically” refers to application of thecompositions of the present invention to the surface of the skin andmucosal cells and tissues (e.g., alveolar, buccal, lingual, masticatory,or nasal mucosa, and other tissues and cells that line hollow organs orbody cavities).

As used herein, the term “pharmaceutically acceptable carrier” refers toany of the standard pharmaceutical carriers including, but not limitedto, phosphate buffered saline solution, water, emulsions (e.g., such asan oil/water or water/oil emulsions), and various types of wettingagents, any and all solvents, dispersion media, coatings, sodium laurylsulfate, isotonic and absorption delaying agents, disintrigrants (e.g.,potato starch or sodium starch glycolate), and the like. Thecompositions also can include stabilizers and preservatives. Forexamples of carriers, stabilizers and adjuvants. (See e.g., Martin,Remington's Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton,Pa. (1975), incorporated herein by reference).

As used herein, the term “pharmaceutically acceptable salt” refers toany salt (e.g., obtained by reaction with an acid or a base) of acompound of the present invention (e.g., in Formulas I-VII) that isphysiologically tolerated in the target subject (e.g., a mammaliansubject, and/or in vivo or ex vivo, cells, tissues, or organs). “Salts”of the compounds of the present invention may be derived from inorganicor organic acids and bases. Examples of acids include, but are notlimited to, hydrochloric, hydrobromic, sulfuric, nitric, perchloric,fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic,toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic,ethanesulfonic, formic, benzoic, malonic, sulfonic,naphthalene-2-sulfonic, benzenesulfonic acid, and the like. Other acids,such as oxalic, while not in themselves pharmaceutically acceptable, maybe employed in the preparation of salts useful as intermediates inobtaining the compounds of the invention and their pharmaceuticallyacceptable acid addition salts. Examples of bases include, but are notlimited to, alkali metal (e.g., sodium) hydroxides, alkaline earth metal(e.g., magnesium) hydroxides, ammonia, and compounds of formula NW₄ ⁺,wherein W is C₁₋₄ alkyl, and the like. Examples of salts include, butare not limited to: acetate, adipate, alginate, aspartate, benzoate,benzenesulfonate, bisulfate, butyrate, citrate, camphorate,camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate,ethanesulfonate, fumarate, flucoheptanoate, glycerophosphate,hemisulfate, heptanoate, hexanoate, chloride, bromide, iodide,2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate,2-naphthalenesulfonate, nicotinate, oxalate, palmoate, pectinate,persulfate, phenylpropionate, picrate, pivalate, propionate, succinate,tartrate, thiocyanate, tosylate, undecanoate, and the like. Otherexamples of salts include anions of the compounds of the presentinvention compounded with a suitable cation such as Na⁺, NH₄ ⁺, and NW₄⁺ (wherein W is a C₁₋₄ alkyl group), and the like. For therapeutic use,salts of the compounds of the present invention are contemplated asbeing pharmaceutically acceptable. However, salts of acids and basesthat are non-pharmaceutically acceptable may also find use, for example,in the preparation or purification of a pharmaceutically acceptablecompound.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to compounds that are modulators ofIntegrin Linked Kinase (ILK), and methods of treating diseases with suchcompounds. In certain embodiments, the compounds are within FormulasI-VII (e.g., Csbl-1). In some embodiments, the compounds are used totreat an ILK-mediated disease, such as cancer (e.g., triple negativebreast cancer) or an inflammatory disease.

Work conducted during the development of embodiments described hereinemployed a structure-based rational design to design and synthesize aseries of small molecule inhibitors to specifically block the Mg-ATPbinding to ILK—a distinct pseudokinase essential for controllingcytoskeleton-dependent cell adhesion and survival in many physiologicaland pathological processes. One compound that was synthesized Csbl-1(Compound 1) was found to inhibit growth of triple negative breastcancer (TNBC) cells and tumors where ILK is highly elevated with noapparent toxicity to healthy breast cells and mice under the sameexperimental conditions. Compounds provided herein are ILK inhibitorsthat are specific for ILK but with little effect to (e.g., do not bindto) EGFR and other structurally similar kinases, and therefore could beless toxic and have a potential to reduce side effects in diseasetreatment.

ILK is a central component of focal adhesions, supramolecular complexesthat link extracellular matrix to actin filaments, regulatingcytoskeleton organization and diverse cytoskeleton-dependent cellularresponses such as cell adhesion, cell shape change, migration,proliferation, and survival (Legate et al., 2006). Being criticallyinvolved in the progression of cancer, diabetes, kidney failure,inflammation, and cardiovascular diseases, ILK has been widelyrecognized as a therapeutic target (Yoganathan et al., 2002; Hannigan etal., 2005; Schmidmaier and Baumann, 2008; Durbin et al., 2009; Cabodi etal., 2010). However, being originally thought to function as a kinase(Hannigan et al., 1996), ILK was later found to contain an unusualpseudoactive site incapable of performing catalysis (Fukuda et al.,2009, 2011), precluding the conventional kinase activity-based drugdevelopment. Interestingly, despite lacking kinase activity, it wasdiscovered that ILK still binds Mg-ATP at the pseudoactive site (Fukudaet al., 2009, 2011) and is capable of utilizing the bound MgATP totransduce non-catalytic signals for modulating cytoskeleton and celladhesion dynamics (Vaynberg et al., 2018). Given that ILK is highlyupregulated in many disease states (Yoganathan et al., 2002; Hannigan etal., 2005; Schmidmaier and Baumann, 2008; Durbin et al., 2009; Cabodi etal., 2010), the compounds provided herein are useful for targeting ILK,yet in conceptually different way, i.e., rather than inhibiting thenon-existent ILK kinase activity, the compounds may be used to block theMgATP binding to ILK and inhibit the relevant pathway(s) anddysfunctional cellular processes in various diseases. Since ILK has noisoform and is highly concentrated especially in cell adhesion sites,the compounds herein could be highly effective in treating ILK-mediateddiseases.

Currently there is no proven targeted therapy for triple negative breastcancer. Furthermore, because ILK has no isoform, ILK specific compoundsherein (e.g., Csbl-1) may have minimal side effects on treating diseaseswhere ILK is elevated, as compared to many known drugs such as erlotiniband gefitinib that target EGFR as well as homologs and structurallysimilar kinases (Kitagawa et al., 2013).

The present invention provides pharmaceutical compositions which maycomprise one or more forms of a compound of Formula I-VII (e.g.,Compound 1), alone or in combination with at least one other agent, suchas a stabilizing compound, or a chemotherapeutic drug, and may beadministered in any sterile, biocompatible pharmaceutical carrier,including, but not limited to, saline, buffered saline, dextrose, andwater.

The methods of the present invention find use in treating (e.g.,prophylactically or therapeutically) ILK-mediated diseases. A compoundof Formula I-VII (e.g., Compound 1) can be administered to a subject(e.g., a patient) intravenously in a pharmaceutically acceptable carriersuch as physiological saline. Standard methods for intracellulardelivery of compounds can be used (e.g., delivery via liposome). Suchmethods are well known to those of ordinary skill in the art. Theformulations of this invention are useful for parenteral administration,such as intravenous, subcutaneous, intramuscular, and intraperitoneal.

As is well known in the medical arts, dosages for any one subject maydepend upon many factors, including the patient's size, body surfacearea, age, the particular compound to be administered, sex, time androute of administration, general health, and interaction with otherdrugs being concurrently administered.

Accordingly, in some embodiments of the present invention, compositionsand/or formulations comprising a compound of Formula I-VII can beadministered to a subject alone, or in combination with other forms of acompound of Formula I-VII, drugs, small molecules, or in pharmaceuticalcompositions where it is mixed with excipient(s) or otherpharmaceutically acceptable carriers. In one embodiment of the presentinvention, the pharmaceutically acceptable carrier is pharmaceuticallyinert. In another embodiment of the present invention, compositionscomprising a compound of Formula I-VII may be administered alone toindividuals subject to or suffering from a disease or condition (e.g.,breast cancer or an inflammatory disease). Compositions comprising acompound of Formula I-VII may be added to a nutritional drink or food(e.g., ENSURE, POWERBAR, or the like), a multivitamin, nutritionalproducts, food products, etc. for daily consumption.

Depending on the target sought to be altered by treatment, thesepharmaceutical compositions may be formulated and administeredsystemically or locally. Techniques for formulation and administrationmay be found in the latest edition of “Remington's PharmaceuticalSciences” (Mack Publishing Co, Easton Pa.). Suitable routes may, forexample, include oral or transmucosal administration; as well asparenteral delivery, including intramuscular, subcutaneous,intramedullary, intrathecal, intraventricular, intravenous,intraperitoneal, or intranasal administration.

For injection, the pharmaceutical compositions of the invention may beformulated in aqueous solutions, preferably in physiologicallycompatible buffers such as Hanks' solution, Ringer's solution, orphysiologically buffered saline. For tissue or cellular administration,penetrants appropriate to the particular barrier to be permeated areused in the formulation. Such penetrants are generally known in the art.

In other embodiments, the pharmaceutical compositions of the presentinvention can be formulated using pharmaceutically acceptable carrierswell known in the art in dosages suitable for oral administration. Suchcarriers enable the pharmaceutical compositions to be formulated astablets, pills, capsules, liquids, gels, syrups, slurries, suspensionsand the like, for oral or nasal ingestion by a patient to be treated.

Pharmaceutical compositions suitable for use in the present inventioninclude compositions wherein the active ingredients are contained in aneffective amount to achieve the intended purpose. For example, aneffective amount of the pharmaceutical agent may be that amount thatalters the expression of a specific gene. Determination of effectiveamounts is well within the capability of those skilled in the art,especially in light of the disclosure provided herein. In addition tothe active ingredients these pharmaceutical compositions may containsuitable pharmaceutically acceptable carriers comprising excipients andauxiliaries which facilitate processing of the active compounds intopreparations which can be used pharmaceutically. The preparationsformulated for oral administration may be in the form of tablets,dragees, capsules, or solutions.

The pharmaceutical compositions of the present invention may bemanufactured in a manner that is itself known (e.g., by means ofconventional mixing, dissolving, granulating, dragee-making, levigating,emulsifying, encapsulating, entrapping or lyophilizing processes).Pharmaceutical formulations for parenteral administration includeaqueous solutions of the active compounds in water-soluble form.Additionally, suspensions of the active compounds may be prepared asappropriate oily injection suspensions. Suitable lipophilic solvents orvehicles include fatty oils such as sesame oil, or synthetic fatty acidesters, such as ethyl oleate or triglycerides, or liposomes. Aqueousinjection suspensions may contain substances which increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol, or dextran. Optionally, the suspension may also containsuitable stabilizers or agents which increase the solubility of thecompounds to allow for the preparation of highly concentrated solutions.

Pharmaceutical preparations for oral use can be obtained by combiningthe active compounds with solid excipient, optionally grinding aresulting mixture, and processing the mixture of granules, after addingsuitable auxiliaries, if desired, to obtain tablets or dragee cores.Suitable excipients are carbohydrate or protein fillers such as sugars,including lactose, sucrose, mannitol, or sorbitol; starch from com,wheat, rice, potato, etc.; cellulose such as methyl cellulose,hydroxypropylmethyl-cellulose, or sodium carboxymethylcellulose; andgums including arabic and tragacanth; and proteins such as gelatin andcollagen. If desired, disintegrating or solubilizing agents may beadded, such as the cross-linked polyvinyl pyrrolidone, agar, alginicacid or a salt thereof such as sodium alginate.

Dragee cores are provided with suitable coatings such as concentratedsugar solutions, which may also contain gum arabic, talc,polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/or titaniumdioxide, lacquer solutions, and suitable organic solvents or solventmixtures. Dyestuffs or pigments may be added to the tablets or drageecoatings for product identification or to characterize the quantity ofactive compound, (i.e., dosage).

Pharmaceutical preparations which can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a coating such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients mixed with a filler orbinders such as lactose or starches, lubricants such as talc ormagnesium stearate, and, optionally, stabilizers. In soft capsules, theactive compounds may be dissolved or suspended in suitable liquids, suchas fatty oils, liquid paraffin, or liquid polyethylene glycol with orwithout stabilizers.

Compositions comprising a compound of the invention formulated in apharmaceutical acceptable carrier may be prepared, placed in anappropriate container, and labeled for treatment of an indicatedcondition. For compositions or formulations comprising a compound ofFormula I-VII, conditions indicated on the label may include treatmentof condition related to prophylactic or therapeutic treatment of canceror an inflammatory disease.

The pharmaceutical composition may be provided as a salt and can beformed with many acids, including but not limited to hydrochloric,sulfuric, acetic, lactic, tartaric, malic, succinic, etc. Salts tend tobe more soluble in aqueous or other protonic solvents that are thecorresponding free base forms. In other cases, the preferred preparationmay be a lyophilized powder in 1 mM-50 mM histidine, 0.1%-2% sucrose,2%-7% mannitol at a pH range of 4.5 to 5.5 that is combined with bufferprior to use.

For any compound used in the methods of the invention, thetherapeutically effective dose can be estimated initially from cellculture assays. Then, preferably, dosage can be formulated in animalmodels (particularly murine models) to achieve a desirable circulatingconcentration range. A therapeutically effective dose refers to thatamount of which ameliorates or prevents symptoms of a disease state orcondition (e.g., through altering gene expression) Toxicity andtherapeutic efficacy of such compounds can be determined by standardpharmaceutical procedures in cell cultures or experimental animals,e.g., for determining the LD₅₀ (the dose lethal to 50% of thepopulation) and the ED₅₀ (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index, and it can be expressed as the ratio LD₅₀/ED₅₀.Compounds which exhibit large therapeutic indices are preferred. Thedata obtained from these cell culture assays and additional animalstudies can be used in formulating a range of dosage for human use. Thedosage of such compounds lies preferably within a range of circulatingconcentrations that include the ED₅₀ with little or no toxicity. Thedosage varies within this range depending upon the dosage form employed,sensitivity of the patient, and the route of administration.

The exact dosage may be chosen by a subject or by a physician in view ofthe patient to be treated. Dosage and administration are adjusted toprovide sufficient levels of the active moiety or to maintain thedesired effect (e.g., alteration of gene expression in a subject).Additional factors that may be taken into account include the severityof the disease state; age, weight, and gender of the patient; diet, timeand frequency of administration, drug combination(s), reactionsensitivities, and tolerance/response to therapy. Long actingpharmaceutical compositions might be administered every 3 to 4 days,every week, or once every two weeks depending on half-life and clearancerate of the particular formulation. In some embodiments, a compound ofFormula I-VII is administered at a daily dose of between 25-600 mg perday (e.g., administered to a subject in such a way so as to providebetween 25-600 mg of a compound of Formula I-VII to the subject eachday). Doses outside of 25-600 mg may be used. In some embodiments, asingle dose of a compound of Formula I-VII is administered once daily.In other embodiments, 2, 3, 4, or more doses may be administered eachday (e.g., once in the morning and once at night, or once every 4 to 6hours). For example, in some embodiments, a compound of Formula I-VII isadministered to a subject in three separate, more than three separate,two separate, or less than two separate doses. In some embodiments, thedaily dose is administered in a time release capsule.

The pharmaceutical compositions of the present invention may beadministered in a number of ways depending upon whether local orsystemic treatment is desired and upon the area to be treated.Administration may be topical (including ophthalmic and to mucousmembranes including vaginal and rectal delivery), pulmonary (e.g., byinhalation or insufflation of powders or aerosols, including bynebulizer; intratracheal, intranasal, epidermal and transdermal), oralor parenteral. Parenteral administration includes intravenous,intraarterial, subcutaneous, intraperitoneal or intramuscular injectionor infusion; or intracranial, e.g., intrathecal or intraventricular,administration.

Pharmaceutical compositions and formulations for topical administrationmay include transdermal patches, ointments, lotions, creams, gels,drops, suppositories, sprays, liquids and powders. Conventionalpharmaceutical carriers, aqueous, powder or oily bases, thickeners andthe like may be necessary or desirable.

Compositions and formulations for oral administration include powders orgranules, suspensions or solutions in water or non-aqueous media,capsules, sachets or tablets. Thickeners, flavoring agents, diluents,emulsifiers, dispersing aids or binders may be desirable.

Compositions and formulations for parenteral, intrathecal orintraventricular administration may include sterile aqueous solutionsthat may also contain buffers, diluents and other suitable additivessuch as, but not limited to, penetration enhancers, carrier compoundsand other pharmaceutically acceptable carriers or excipients.

Thus, in some embodiments, pharmaceutical compositions of the presentinvention include, but are not limited to, solutions, emulsions, andliposome-containing formulations. These compositions may be generatedfrom a variety of components that include, but are not limited to,preformed liquids, self-emulsifying solids and self-emulsifyingsemisolids. The pharmaceutical formulations of the present invention,which may conveniently be presented in unit dosage form, may be preparedaccording to conventional techniques well known in the pharmaceuticalindustry. Such techniques include the step of bringing into associationthe active ingredients with the pharmaceutical carrier(s) orexcipient(s). In general the formulations are prepared by uniformly andintimately bringing into association the active ingredients with liquidcarriers or finely divided solid carriers or both, and then, ifnecessary, shaping the product.

Thus, in some embodiments, the compositions of the present invention maybe formulated into any of many possible dosage forms such as, but notlimited to, tablets, capsules, liquid syrups, soft gels, suppositories,and enemas. The compositions of the present invention may also beformulated as suspensions in aqueous, non-aqueous or mixed media.Aqueous suspensions may further contain substances that increase theviscosity of the suspension including, for example, sodiumcarboxymethylcellulose, sorbitol and/or dextran. The suspension may alsocontain stabilizers.

In one embodiment of the present invention the pharmaceuticalcompositions may be formulated and used as foams. Pharmaceutical foamsinclude formulations such as, but not limited to, emulsions,microemulsions, creams, jellies and liposomes. While basically similarin nature these formulations vary in the components and the consistencyof the final product. The compositions of the present invention mayadditionally contain other adjunct components conventionally found inpharmaceutical compositions. Thus, for example, the compositions maycontain additional, compatible, pharmaceutically-active materials suchas, for example, antipruritics, astringents, local anesthetics oranti-inflammatory agents, or may contain additional materials useful inphysically formulating various dosage forms of the compositions of thepresent invention, such as dyes, flavoring agents, preservatives,antioxidants, opacifiers, thickening agents and stabilizers. However,such materials, when added, should not unduly interfere with thebiological activities of the components of the compositions of thepresent invention. The formulations can be sterilized and, if desired,mixed with auxiliary agents, e.g., lubricants, preservatives,stabilizers, wetting agents, emulsifiers, salts for influencing osmoticpressure, buffers, colorings, flavorings and/or aromatic substances andthe like which do not deleteriously interact with the nucleic acid(s) ofthe formulation. In some embodiments, the invention providespharmaceutical compositions containing (a) one or more forms of acompound of Formula I-VII and (b) one or more other agents (e.g.,anti-cancer therapeutic).

The present invention also includes methods involving co-administrationof compounds comprising a compound of Formula I-VII described hereinwith one or more additional active agents. Indeed, it is a furtheraspect of this invention to provide methods for enhancing prior arttherapies and/or pharmaceutical compositions by co-administering acomposition comprising a compound of Formula I-VII of this invention. Inco-administration procedures, the agents may be administeredconcurrently or sequentially. In one embodiment, the compounds describedherein are administered prior to the other active agent(s). Thepharmaceutical formulations and modes of administration may be any ofthose described above. In addition, the two or more co-administeredagents may each be administered using different modes or differentformulations.

In certain embodiments, the compounds described herein are administeredto a subject with an ILK-mediated disease or condition. Examples of suchdiseases and conditions include, but are not limited to, such diseasesand conditions described in the literature. One example of such adisease is cancer, including, but not limited to, leukemia (see, e.g.,de la Puente et al., Leuk Res. 2015 Sep. 9. pii: S0145-2126(15)30377-5,herein incorporated by reference), colon cancer (see, e.g., Tsoumas etal., Cancer Genomics Proteomics. 2018 March-April; 15(2):127-141; andBravou et al., J Pathol. 2006 January; 208(1):91-9; and Marotta et al.,Br J Cancer. 2003 Jun. 2; 88(11):1755-62; all of which are hereinincorporated by reference), lung cancer (see, e.g., Karachaliou et al.,EBioMedicine, 2018 Nov. 22. pii: S2352-3964(18)30541-3, hereinincorporated by reference), prostate cancer (see, e.g., Hu et al., OncolLett. 2018 October; 16(4):4945-4952, herein incorporated by reference),pancreatic cancer (see, e.g., Schaeffer et al., Virchows Arch. 2010March; 456(3):261-8, herein incorporated by reference), kidney cancer(see, e.g., Han et al., Mol Cancer her. 2015 April; 14(4):1024-34; andAgouni et al., Carcinogenesis. 2007 September; 28(9):1893-901; both ofwhich are herein incorporated by reference); glioblastoma cancer (see,e.g., Hausmann et al., Oncotarget. 2015 Oct. 27; 6(33):34592-605; andVerano-Braga et al., Oncotarget. 2018 Jan. 25; 9(10):9400-9414; both ofwhich are herein incorporated by reference); breast cancer (see, e.g.,Qu et al., Mol Med Rep. 2017 October; 16(4):5036-5042; and Pang et al.,Cancer Res. 2016 Sep. 15; 76(18):5277-87; both of which are hereinincorporated by reference); myeloma (see, e.g., Zhao et al., Oncol Lett.2018 July; 16(1):1101-1106; herein incorporated by reference), and skincancer (see, e.g., Dai et al., Clin. Cancer Res. 2003 Oct. 1;9(12):4409-14 (herein incorporated by reference). Other examples of sucha diseases and conditions include, but are not limited to, diabetes(see, e.g., Huang et al., Can J Diabetes. 2017 June; 41(3):312-321;Williams et al., Diabetes. 2017 February; 66(2):325-334; and Kang etal., Diabetes. 2016 June; 65(6):1590-600; all of which are hereinincorporated by reference), thrombosis (see, e.g., Jones et al., JThromb Haemost. 2014 August; 12(8):1342-52, herein incorporated byreference), aging (see, e.g., Nishimura et al., Aging Cell. 2014 June;13(3):431-40; herein incorporated by reference), heart diseases (e.g.,Traister et al., PLoS One. 2012; 7(5):e37802; Sopko et al., PLoS One.2011; 6(5):e20184; and Lu et al., Circulation. 2006 Nov. 21;114(21):2271-9; all of which are herein incorporated by reference),pulmonary disease (see, e.g., Wu et al., Am J Physiol Lung Cell MolPhysiol. 2008 December; 295(6):L988-97; and Kudryashova et al., Am JRespir Crit Care Med. 2016 Oct. 1; 194(7):866-877; both of which areherein incorporated by reference).

EXPERIMENTAL

The following examples are provided in order to demonstrate and furtherillustrate certain preferred embodiments and aspects of the presentinvention and are not to be construed as limiting the scope thereof.

Example 1 ILK Inhibitor Development and Characterization

This Example describes the development and characterization of ILKinhibitors.

Development and Biochemical Characterization of Novel ILK Inhibitors

To develop a specific ILK inhibitor, we performed computer-aideddocking-based virtual screening on the Mg-ATP-bound pseudoactive site ofILK against a public database (NCI Plated 2007 and NCI Diversity 3)containing ˜0.1 million drug-like molecules. However, none of thecompounds ranked on the top list showed any detectable binding topurified ILK using fluorescence-based method. We then examined severalknown ILK inhibitors, which include QLT-0267 (Troussard et al., 2006),compound 22 or T315 (Lee et al., 2011), chelidonine (Kim et al., 2015),which were identified by cell-based assays. Surprisingly, thesecompounds also had no detectable binding to purified ILK despitepreviously suggested activity.

Thus, while these compounds were extensively used in studyingILK-mediated cellular and disease processes, their reported inhibitoryeffects are probably due to unknown artifacts or indirect bindingevents. Next, we turned our attention to previously reported studies onkinase profiling and quantitative chemical proteomics (Conradt et al.,2011; Patricelli et al., 2011; Augustin et al., 2013). These studiessuggested that a widely known lung cancer drug erlotinib, which targetsEGFR, might also bind to ILK as an off-target (Conradt et al., 2011;Patricelli et al., 2011; Augustin et al., 2013). By performing a robustfluorescence-based binding assay, we found that the FDA approved drugErlotinib (TARCEVA) indeed binds potently to purified recombinant ILK atK_(D)˜0.43M, which is very close to the affinity of Erlotinib to EGFRmeasured at the same experimental conditions (K_(D)˜0.31 μM) (Table 1).Another erlotinib-like EGFR inhibitor Gefitinib exhibited 10-fold weakerbinding affinity to ILK (K_(D)˜4.51 μM) yet ˜3-fold stronger affinity toEGFR (K_(D)˜0.11 μM) than erlotinib (Table 1). To understand thestructural basis of the erlotinib/gefitinib binding to ILK, wedetermined high resolution crystal structures of erlotinib/ILK andgefitinib/ILK complexes, which revealed similar drug binding modes butwith erlotinib having more hydrophobic contacts to ILK than gefitinib,explaining the reason behind strong binding of erlotinib to ILK comparedto gefitinib (FIG. 1 panel B).

The findings above have propelled us to explore the possibility ofdeveloping specific ILK inhibitors using the high-resolution structureof erlotinib/ILK complex vs known EGFR/erlotinib structure (PDB code1M17). Our rationale was to use these structures to guide the design ofa novel and selective compound that favors ILK but disfavors EGFR.

Detailed structural comparison between the erlotinib/ILK complex and theerlotinib/EGFR complex (FIG. 1 panel A) revealed unique features of theformer, which allowed us to design numerous compounds (Table 1) thathave been subjected to binding studies. Of these compounds, Csbl-1(Compound 1) binds to the pseudoactive site of ILK but unfavorably toEGFR (Table 1).

TABLE 1 Binding ILK - EGFR - Difference Binding Binding Binding ILK Ex-affinity affinity (Kd)/EGFR Name Structure periment (nM) (nM) (Kd)Erlotinib

EnSpire fluores- cence intensity 430 +/− 50 310 +/− 30 1.3870968Gefitinib

EnSpire fluores- cence intensity 4510 +/− 720 110 +/− 10 41 Erlotinib-4-methylphenyl analog (E4ME)

EnSpire fluores- cence intensity 1546 +/− 222 745 +/− 74.8 2.075168Compound 1 (Csbl-1)

EnSpire fluores- cence intensity 2770 +/− 290 17,700 +/− 1700 0.156497Compound 3 (cmpd#3)

EnSpire fluores- cence intensity 49,570 +/− 2210 N/A N/A Compound 4(cmpd#4)

EnSpire fluores- cence intensity 1545 +/− 176 1153 +/− 161.4 1.3399827Compound 1 New (cmpd#1new)

Nano- temper 6852.8 +/− 607.27 12,658 +/− 1970.9 0.5413809 Compound 2New (cmpd#2new)

Nano- temper 7995 +/− 527 N/A N/A Compound 7 (cmpd #7)

Nano- temper 2444.6 +/− 149.08 6588.4 +/− 464.84 0.371046 Compound 8(Csbl-2)

Nano- temper 922.12 +/− 49.959 8840 +/− 671 0.104312 Compound 22

EnSpire fluores- cence intensity N/D N/D N/D QLT0267-like compound(QLT0267)

EnSpire fluores- cence intensity N/D N/D N/D Chelidonine

EnSpire fluores- cence intensity N/D N/D N/D

Csbl-1 retains quinazoline feature of erlotinib but is chemicallydistinct with totally different structure (Table 1). The compound isalso totally different from previously reported ILK inhibitors that didnot bind to purified ILK in our experiments (see Table 1).

To experimentally verify the effectiveness of our design, we synthesizedCsbl-1 (Compound 1) and performed the affinity measurement. As expected,Csbl-1 binds potently to ILK at K_(D)˜2.77 μM (Table 1) yet unfavorablyto EGFR with an affinity that is too weak to be accurately determinedunder the same experimental condition. Consistently, Csbl-1 has adramatically reduced IC₅₀ (77.2 μM) to EGFR vs erlotinib (1.5 μM) (>50times reduction). Thus, being potent to bind ILK, Csbl-1 is expected toexert minimal effect on EGFR (e.g., when properly dosed). We alsodesigned Csbl-2 (Compound 8), a Csbl-1 variant, which binds ˜3-foldtighter to ILK (K_(D)˜0.92 μM (Table 1) than Csbl-1 to ILK (K_(D)˜2.77μM) (Table 1). However, Csbl-2 still binds to EGFR at the K_(D)˜8.84 μM.In other words, while Csbl-2 has stronger affinity to ILK than Csbl-1,Csbl-2 might exhibit stronger off-target effect to EGFR than Csbl-1depending on the drug dosage and disease conditions.

It is important to note here that drug binding affinity or IC₅₀ may varysignificantly depending on the measurement methods and experimentalconditions. For example, the IC₅₀ for erlotinib to inhibit purified EGFRis 1.51 μM in our kinase assay using purified EGFR, which is 75 timeslower than the previously reported ICs-20 nM based on a cell-based assay(Moyer et al., 1997). Thus, comparison of the binding affinities or IC₅₀is only meaningful under the same experimental conditions. Under thesame experimental conditions, the affinity of Csbl-1 to ILK (K_(D)˜2.77μM) is ˜6 times weaker than that of erlotinib to ILK (K_(D)˜0.43 μM)(Table 1). However, Csbl-1 is much more selective to ILK than erlotinibwith the latter almost having no selectivity to ILK and EGFR (Table1).Furthermore, since ILK is extremely abundant in cells (˜20 μM, ˜12% ofactin in non-muscle cells) (Pollard et al., 2000; Zeiler et al., 2014)and its local concentration, e.g., in FAs, is even much higher(likely >submM), it is proposed that Csbl-1 would be quite potent andselective to inhibit the ILK pathway(s) in cells or in diseaseconditions where ILK level is further elevated. By contrast, EGFR, whichhas extremely low level even in cancer patients (<35 nM)(Jantus-Lewintre et al., 2011), is expected to be generally insensitiveto this inhibitor.

Example 2 Characterization of Csbl-1

This Example describes further characterization of Csbl-1 (Compound 1).

Crystal Structure of Csbl-1 Bound to ILK Reveals a Basis for theInhibitor Specificity

To elucidate definitively how Csbl-1 specifically binds to ILK, wedetermined the crystal structure of Csbl-1/ILK complex. FIG. 2 panel Ashows that the Csbl-1 binding would clearly occlude the Mg-ATP bindingto ILK. Superposition of Csbl-1/ILK and erlotinib/EGFR structuresfurther reveals that Csbl-1 would indeed have unfavorable binding toactive EGFR, e.g., exerting steric clashes with a conserved bulky F778in Gly rich P loop (corresponding to S204 in ILK) (FIG. 2 panel B). Thusour structure provides a hypothetical atomic basis for understanding whyCsbl-1 binds selectively to ILK but unfavorably to EGFR (Table 1). Acomprehensive kinase inhibitor off-target analysis suggested thaterlotinib may bind more than a dozen of off target kinases with >60%inhibition efficiency (Kitagawa et al., 2013). Analysis of the crystalstructures of these kinases or their homologs reveals that they sharemost of the key structural features such as activation loopconfiguration and conserved bulky Phe in the Gly rich P loop as seen inactive EGFR, which will cause similar steric clashes with Csbl-1. Thus,Csbl-1 is highly selective to ILK pseudoactive site that is distinctfrom these EGFR-like kinases. Consistently, a proteomic analysis ofCsbl-1 against all known FDA-approved kinase inhibitors revealed thatCsbl-1 has minimal off-target effect to kinases (FIG. 3 ). Csbl-1(compound 1) also exhibits excellent pharmacological ADME (Absorption,Distribution, Metabolism, and Excretion) properties such as toxicity,plasma stability, etc. as shown in Tables 2-12 below.

TABLE 2 Permeability Results of Test Compounds in Caco2 Cell LineP_(app (A-B)) P_(app (B-A)) Efflux Recovery Recovery Compound ID (10⁻⁶,cm/s) (10⁻⁶, cm/s) Ratio (%)AP-BL (%)BL-AP Propranolol 19.46 15.88 0.8260.83 80.03 Digoxin 0.61 16.59 27.01 87.87 96.54 Prazosin 12.46 30.092.41 85.14 99.05 Compound 1 15.38 7.31 0.48 50.97 53.50

TABLE 3 The Assessment of Caco2 Cell Monolayer Integrity LY LYTEER_(A-B) TEER_(B-A) Leakage_(A-B) Leakage_(B-A) Compound ID (Ω · cm²)(Ω · cm²) (%) (%) Propranolol 353 349 0.32 0.29 Digoxin 520 484 0.230.20 Prazosin 551 556 0.17 0.26 Compound 1 529 547 0.24 0.18

TABLE 4 Stability results of Test Compound and control compound in PBSat pH 7.4 and SIF Remaining Percentage (%) Compound PH Value 0 min 30min 60 min 120 min 180 min 240 min Chlorambucil 7.4 100.00 37.17 11.532.35 0.09 0.03 Compound 1 7.4 100.00 102.02 105.41 103.04 104.93 99.54Chlorambucil SIF 100.00 58.85 33.12 8.26 2.57 0.79 Compound 1 SIF 100.00104.86 109.34 105.85 108.93 109.26

TABLE 5 Inhibition of 9 CYP Isoforms by Test Compounds in Human LiverMicrosomes IC₅₀ (μM) Compound ID 1A2 2A6 2B6 2C8 2C9 2C19 2D6 2E1 3A4-MFurafylline 2.19 — — — — — — — — Tranylcypromine — 0.69 — — — — — — —Sulfaphenazole — — — — 0.66 — — — — N-3-benzylnirvanol — — — — — 0.25 —— — Quinidine — — — 3.72 — — 0.11 — — Ketoconazole — — 1.46 — — — — —0.018 Quercetin — — — — — — — — — Disulfiram — — — — — — — 37.71 —Compound 1 33.29 >50 >50 3.24 13.82 10.77 27.57 >50 43.78

TABLE 6 Metabolic Stability of Test Compounds in Liver Microsomes ofDifferent Species (a) CL_(int) Scaled- T_(1/2) (μL/min/mg up CL_(int)Compound ID Species (min) protein) (mL/min/Kg) Verapamil Human 13.32104.07 130.53 Rat 5.54 250.12 448.21 Mouse 11.56 119.86 524.39 Compound1 Human 34.81 39.82 49.94 Rat 16.00 86.61 155.20 Mouse 35.36 39.20171.50

TABLE 7 Metabolic Stability of Test Compounds in Liver Microsomes ofDifferent Species (b) Remaining Percentage (%) Compound ID Species AssayFormat 0 min 15 min 30 min 45 min 60 min Verapamil Human With NADPH100.00 26.29 10.84 5.95 4.30 Without NADPH 100.00 88.70 84.00 83.5090.40 Rat With NADPH 100.00 12.23 2.35 0.71 0.40 Without NADPH 100.00102.19 92.87 91.71 97.12 Mouse With NADPH 100.00 15.30 6.34 3.32 2.42Without NADPH 100.00 106.23 89.98 93.40 111.61 Compound 1 Human WithNADPH 100.00 70.61 51.77 38.83 30.40 Without NADPH 100.00 98.20 92.8189.22 103.59 Rat With NADPH 100.00 45.00 21.08 11.69 7.65 Without NADPH100.00 95.03 90.68 91.30 102.48 Mouse With NADPH 100.00 72.40 51.6737.39 32.04 Without NADPH 100.00 93.90 93.90 91.46 94.51

TABLE 8 Log D Results of Test Compound in 1-Octanol/PBS pH 7.4 CompoundID LogD Value Progesterone 3.78 Compound 1 3.61

TABLE 9 Mitochondrial toxicity of the test compound in HepG2 cells FoldIC₅₀ IC₅₀ change Compound ID (μM)_Glucose (μM)_Galactose in IC₅₀Compound 1 >150 >150 — Nefazodone 77.22 6.82 11.33 Digitonin 4.30 1.792.40 * Compound precipitation observed for Compound 1 at 50 and 150 μM.

TABLE 10 Protein Binding Results of Test Compound in Human, Rat andMouse Plasma Human Plasma Rat Plasma Mouse Plasma Compound ID % Bound %Recovery % Bound % Recovery % Bound % Recovery Ketoconazole 98.33 101.0199.63 100.83 99.33 104.25 Compound 1 96.97 98.77 99.17 93.40 99.00109.72

TABLE 11 Stability results of Compound 1 and control compounds in human,rat and mouse plasma Remaining Percentage (%) Compound Species 0 min 15min 30 min 45 min 60 min 120 min Propantheline Human 100.00 54.01 26.589.84 3.10 0.13 Compound 1 Human 100.00 93.79 93.79 90.52 90.20 88.56Mevinolin Rat 100.00 3.78 0.00 0.00 0.00 0.00 Compound 1 Rat 100.0092.93 94.35 93.29 94.70 91.52 Propantheline Mouse 100.00 74.15 64.0348.07 37.61 14.60 Compound 1 Mouse 100.00 96.17 96.17 97.56 96.17 93.73

TABLE 12 Solubility Results of Test Compound and Control Compound in PBSat pH 7.4 Compound ID Solubility (μM) Progesterone 13.88 Compound 1 7.96Csbl-1 Impairs ILK-PINCH-Parvin (IPP)-Mediated Actin Bundling In Vitro

Next, we set out to obtain in vitro evidence of Csbl-1. It was recentlyshowed that ILK-centered IPP complex promotes actin filament bundling(Vaynberg et al., 2017), which is mediated by two actin binding motifsin IPP as well as MgATP at the ILK pseudoactive site. Mutation of L207into bulky Trp in ILK abolished the MgATP binding to ILK and impairedthe formation and morphology of actin bundles (Vaynberg et al., 2018).Using the same actin bundling assay, we examined the effect of Csbl-1 onthe IPP-mediated actin bundling. FIG. 4 shows that Csbl-1 caused thesubstantial change of IPP-induced actin bundles. In general, morebundles with smaller sizes were observed as a result of Csbl-1interference. Csbl-1 would knock out MgATP at the ILK site and increasethe local concentration of Mg²⁺, which may impact the actin bundlingprocess (Tang and Janmey, 1996, Vaynberg et al., 2018). These dataprovide evidence for the effect of Csbl-1 in vitro, i.e., it altersILK-mediated actin filament assembly.

Csbl-1 Inhibits ILK-Mediated Proliferation of Triple Negative BreastCancer (TNBC) Cell but not Normal Breast Cell

Since ILK-dependent actin assembly regulates actin stress fibers, whichin turn affects many cellular processes such as cell spreading,proliferation, and survival, we expected that Csbl-1 would interferewith ILK-dependent dysfunctional cellular behaviors. We thereforedecided to examine the effect of Csbl-1 on MDA-MB-231— a triple negativebreast cancer (TNBC) cell line. This cell line was chosen for thefollowing reasons: (i) ILK is highly elevated in this metastatic cellline (see, FIG. 5 a in Mongroo et al., 2004; FIG. 1E in Hsu et al.,2015; FIG. 1A in Qu et al., 2017)); (ii) it is a major type of TNBC cellline and has been extensively characterized (Holiday et al., 2011);(iii) currently there is no proven targeted therapy for TNBC (Denkert etal., 2017) so any effect of Csbl-1 would facilitate the development oftherapeutics to treat this devastating disease. FIG. 5 panel A showsthat Csbl-1 had little effect on MCF10a—non-tumorigenic epithelialbreast cells whereas erlotinib exhibited significant toxicity to thesecells probably due to off-target effects (Patricelli et al., 2011). Bycontrast, Csbl-1 substantially inhibited the proliferation of MDA-MB-231whereas erlotinib had little effect at the same dose (FIG. 5 panel B). Adose-dependent titration indicated that the IC₅₀ of Csbl-1 on MDA-MB-231is ˜190 nM. These data provide strong evidence that Csbl-1 is specificto inhibit TNBC cells.

Csbl-1 Inhibits TNBC Tumor Growth Based on a Xenograft Model

The next step was to evaluate the effect of Csbl-1 in mice bearing TNBCtumors. Given the fact that higher ILK expression level is correlatedwith poor outcome and reduced survival for TNBC patients (FIG. 6 ) andthe above TNBC cell-based data, we decided to test whether Csbl-1 wouldbe effective in suppressing the TNBC tumors in mice. Using Csbl-1dissolved in CMC/TWEEN 80, normal mice were orally fed with 25 mg/kg, 50mg/kg, 75 mg/kg, 100 mg/kg, 200 mg/kg, 400 mg/kg, and 510 mg/kg dosesevery day for two weeks and the mice were all fine with no toxicsymptoms. Xenograft TNBC tumors were generated using MDA-MB231 on 8mice. At 10¹ day when the tumors were ˜3 mm, 4 mice were fed with Csbl-1at 100 mg/kg five times/week and the other 4 mice (control, no drug) hadno Csbl-1 treatment. With no significant weight difference between thecontrol mice fed with DMSO and mice fed with the compound Csbl-1. FIG. 7shows that Csbl-1 significantly suppressed the TNBC tumor growth by ˜40%at metastatic phase as compared to DMSO, which is consistent with thecell-based data in FIG. 5 . We also examined the Csbl-1 effect on themouse models with human derived patient TNBC tumor (PDX). FIG. 8 showsthat Csbl-1 effectively inhibited TNBC tumor growth in PDX mice. Thesedata provide in vivo evidence that Csbl-1 is a potent inhibitor of TNBC.

Example 3 Synthesis of Compounds

This Example describes exemplary synthesis protocols that could beemployed to synthesize Csbl-1 (Compound 1) and Csbl-2 (Compound 8).

The synthesis of compounds of Formula I-VII can be achieved by skilledsynthetic chemists using, for example, the procedures and intermediatesexemplified above (see also Schnur and Arnold, 1998; Pandey et al.,2002; Li et al., 2013; Tung, 2015; all of which are herein incorporatedby reference). Additional methods for synthesizing compounds of FormulaI-VII including those within routes not explicitly shown in the aboveschemes are within the means of chemists of ordinary skill in the art.

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All publications and patents mentioned in the above specification areherein incorporated by reference. Various modifications and variationsof the described compositions and methods of the invention will beapparent to those skilled in the art without departing from the scopeand spirit of the invention. Although the invention has been describedin connection with specific preferred embodiments, it should beunderstood that the invention as claimed should not be unduly limited tosuch specific embodiments. Indeed, various modifications of thedescribed modes for carrying out the invention that are obvious to thoseskilled in the relevant fields are intended to be within the scope ofthe present invention.

We claim:
 1. A compound having a structure of Formula I, or apharmaceutically acceptable salt thereof, wherein Formula I is:

wherein R1-R7, R13-R16 and X1 are each an atom or group independentlyselected from: hydrogen, deuterium, trifluoromethyl, lower alkyl, loweralkoxy, aryloxy, trifluoromethoxy, —SCF₃, cyano, nitro, amino, loweralkylamino, lower dialkylamino, mercapto, lower alkylthio, arylthio,formyl, lower alkylcarbonyl, arylcarbonyl, lower alkylcarboxy,arylcarboxy, lower alkoxylcarboxy, aryloxylcarboxy, lower alkanoylamino,arylcarbonylamino, carbamido, lower alkylcarbamido, arylcarbamido,aminocarboxy, lower alkylaminocarboxy, arylaminocarboxy,trifluoroacetyl, halogen, hydroxylcarbonyl, lower alkoxylcarbonyl,aryloxycarbonyl, sulfinyl, lower alkylsulfinyl, arylsulfinyl, sulfonyl,lower alkylsulfonyl, arylsulfonyl, sulfonamido, lower alkylsulfonamido,arylsulfonamido, and aryl; wherein at least one of R1-R7, R14-R19 and X1is not hydrogen; wherein R17-R19 are each independently selected from:hydrogen, deuterium, trifluoromethyl, lower alkyl, lower alkoxy,aryloxy, trifluoromethoxy, —SCF₃, cyano, nitro, amino, lower alkylamino,lower dialkylamino, mercapto, lower alkylthio, arylthio, loweralkylcarbonyl, arylcarbonyl, formyl, lower alkylcarboxy, arylcarboxy,lower alkoxylcarboxy, aryloxylcarboxy, formamido, lower alkanoylamino,arylcarbonylamino, carbamido, lower alkylcarbamido, arylcarbamido,aminocarboxy, lower alkylaminocarboxy, arylaminocarboxy,trifluoroacetyl, halogen, hydroxylcarbonyl, lower alkoxylcarbonyl,aryloxycarbonyl, sulfinyl, lower alkylsulfinyl, arylsulfinyl, sulfonyl,lower alkylsulfonyl, arylsulfonyl, sulfonamido, lower alkylsulfonamido,arylsulfonamido, aryl, —OH, —CH₂—OH, —CH₂—NH₂, —CH₂—NO₂, —CH₂—COOH,—CH₂—CN, and —CH₂—CONH₂; wherein R8-R12 are each independently selectedfrom: hydrogen, deuterium, fluorine, chlorine, bromine, and iodine;wherein X2 is N or CH; wherein X3 is NH, O, S, or CH₂; and wherein X4 isselected from: fluorine, chlorine, bromine, iodine, —CN, and —C≡CY whereY is hydrogen or deuterium.
 2. A compound having a structure of FormulaII, or a pharmaceutically acceptable salt thereof, wherein Formula IIis:

wherein R1-R14, and R20-R23 are each an atom or group independentlyselected from: hydrogen, deuterium, lower alkyl, lower alkoxy, aryloxy,trifluoromethyl, trifluoromethoxy, —SCF₃, cyano, nitro, amino, loweralkylamino, lower dialkylamino, mercapto, lower alkylthio, arylthio,formyl, lower alkylcarbonyl, arylcarbonyl, lower alkylcarboxy,arylcarboxy, lower alkoxylcarboxy, aryloxylcarboxy, formamido, loweralkanoylamino, arylcarbonylamino, carbamido, lower alkylcarbamido,arylcarbamido, aminocarboxy, lower alkylaminocarboxy, arylaminocarboxy,trifluoroacetyl, halogen, hydroxylcarbonyl, lower alkoxylcarbonyl,aryloxycarbonyl, sulfinyl, lower alkylsulfinyl, arylsulfinyl, sulfonyl,lower alkylsulfonyl, arylsulfonyl, sulfonamido, lower alkylsulfonamido,arylsulfonamido, and aryl, and wherein at least one among R1-R14, R21-R2is not hydrogen; wherein R24-R26 are each independently selected from:hydrogen, deuterium, lower alkyl, lower alkoxy, aryloxy,trifluoromethyl, trifluoromethoxy, —SCF₃, cyano, nitro, amino, loweralkylamino, lower dialkylamino, mercapto, lower alkylthio, arylthio,lower alkylcarbonyl, arylcarbonyl, formyl, lower alkylcarboxy,arylcarboxy, lower alkoxylcarboxy, aryloxylcarboxy, formamido, loweralkanoylamino, arylcarbonylamino, carbamido, lower alkylcarbamido,arylcarbamido, aminocarboxy, lower alkylaminocarboxy, arylaminocarboxy,trifluoroacetyl, halogen, hydroxylcarbonyl, lower alkoxylcarbonyl,aryloxycarbonyl, sulfinyl, lower alkylsulfinyl, arylsulfinyl, sulfonyl,lower alkylsulfonyl, arylsulfonyl, sulfonamido, lower alkylsulfonamido,arylsulfonamido, aryl, —OH, —CH₂—OH, —CH₂—NH₂, —CH₂—NO₂, —CH₂—COOH,—CH₂—CN, and —CH₂—CONH₂; wherein R15-R19 are each independently selectedfrom: hydrogen, deuterium, fluorine, chlorine, bromine, and iodine;wherein X1 is nitrogen or CH; wherein X2 is selected from: NH, O, S, andCH₂; and wherein X3 is selected from: fluorine, chlorine, bromine,iodine, —CN, and —C≡CY where Y is hydrogen or deuterium.
 3. A compoundhaving a structure of Formula III, or a pharmaceutically acceptable saltthereof, wherein Formula III is:

wherein X1 is n-propylene, and wherein 1 to 6 hydrogen atoms areoptionally replaced in said n-propylene with deuterium atoms; wherein X2is selected from: —OH, —OD, —OCH₃, —OCH₂D, —OCHD₂, and —OCD₃, wherein Dis deuterium; wherein X3 is N or CH; wherein X4 is selected from: NH, O,S, and CH₂; X5 is selected from: fluorine, chlorine, bromine, iodine,—CN, and —C≡CY where Y is hydrogen or deuterium: wherein R1-R8 are eachindependently selected from: hydrogen and deuterium; wherein R9-R13 areeach independently selected from: hydrogen, deuterium, fluorine,chlorine, bromine, and iodine; wherein R14-R17 are each an atom or groupindependently selected from: hydrogen, deuterium, lower alkyl, loweralkoxy, aryloxy, trifluoromethyl, trifluoromethoxy, —SCF₃, cyano, nitro,amino, lower alkylamino, lower dialkylamino, mercapto, lower alkylthio,arylthio, formyl, lower alkylcarbonyl, arylcarbonyl, lower alkylcarboxy,arylcarboxy, lower alkoxylcarboxy, aryloxylcarboxy, formamido, loweralkanoylamino, arylcarbonylamino, carbamido, lower alkylcarbamido,arylcarbamido, aminocarboxy, lower alkylaminocarboxy, arylaminocarboxy,trifluoroacetyl, halogen, hydroxylcarbonyl, lower alkoxylcarbonyl,aryloxycarbonyl, sulfinyl, lower alkylsulfinyl, arylsulfinyl, sulfonyl,lower alkylsulfonyl, arylsulfonyl, sulfonamido, lower alkylsulfonamido,arylsulfonamido, and aryl, wherein at least one among R14-R20 is nothydrogen; and wherein R18-R20 are each independently selected from:hydrogen, deuterium, lower alkyl, lower alkoxy, aryloxy,trifluoromethyl, trifluoromethoxy, —SCF₃, cyano, nitro, amino, loweralkylamino, lower dialkylamino, mercapto, lower alkylthio, arylthio,formyl, lower alkylcarbonyl, arylcarbonyl, lower alkylcarboxy,arylcarboxy, lower alkoxylcarboxy, aryloxylcarboxy, formamido, loweralkanoylamino, arylcarbonylamino, carbamido, lower alkylcarbamido,arylcarbamido, aminocarboxy, lower alkylaminocarboxy, arylaminocarboxy,trifluoroacetyl, halogen, hydroxylcarbonyl, lower alkoxylcarbonyl,aryloxycarbonyl, sulfinyl, lower alkylsulfinyl, arylsulfinyl, sulfonyl,lower alkylsulfonyl, arylsulfonyl, sulfonamido, lower alkylsulfonamido,arylsulfonamido, aryl, —OH, —CH₂—OH, —CH₂—NH₂, —CH₂—NO₂, —CH₂—COOH,—CH₂—CN, and —CH₂—CONH₂.
 4. A compound selected from the groupconsisting of:


5. A method of treating triple-negative breast cancer in a subject inneed thereof, comprising: administering a compound of claim 1 to thesubject.
 6. The method of claim 1, wherein said subject is a human. 7.The method of claim 5, wherein said compound is co-administered with ananti-cancer agent and/or an anti-inflammatory agent.
 8. The method ofclaim 5, wherein said treating comprises administering between 0.05-3000mg of said compound to said subject.
 9. The method of claim 5, whereinsaid treating comprises administering between 25-600 mg of said compoundto said subject.
 10. The method of claim 5, wherein said treatingcomprises administering 25-600 mg to said subject per day for at leasttwo days.
 11. A method of treating triple-negative breast cancer in asubject in need thereof, comprising: administering a compound of claim 2to the subject.
 12. The method of claim 11, wherein said subject is ahuman.
 13. A method of treating triple-negative breast cancer in asubject in need thereof, comprising: administering a compound of claim 3to the subject.
 14. The method of claim 13, wherein said subject is ahuman.
 15. A method of treating triple-negative breast cancer in asubject in need thereof, comprising: administering a compound of claim 4to the subject.
 16. The method of claim 15, wherein said subject is ahuman.
 17. A pharmaceutical composition comprising a compound of claim 1or a pharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier.
 18. A pharmaceutical composition comprising acompound of claim 2 or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier.
 19. A pharmaceutical compositioncomprising a compound of claim 3 or a pharmaceutically acceptable saltthereof, and a pharmaceutically acceptable carrier.
 20. A pharmaceuticalcomposition comprising a compound of claim 4 or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier.